骨碎补结合组织工程软骨促进软骨再生的实验研究

Experimental Study on Effect of Drynaria Combining with Tissue Engineered Cartilage to Promote Cartilage Regeneration

目的:评估骨碎补结合组织工程软骨治疗对实验兔软骨缺陷模型软骨再生的疗效。方法:将h IGF-1基因转染MSCs,并与脱细胞真皮基质(ADM)构建组织工程软骨。24只新西兰白兔随机分为A、B、C、D四组,A、C组进行自体软骨移植,B、D组进行改建的细胞-ADM移植。C、D组用40%骨碎补汤喂养4周,150 m L/d。第12周处死实验动物,分离缺损关节软骨部位,蜡块包埋染色,通过总体形态评价软骨再生组织。采用组织学评分评估再生软骨质量。采用甲苯胺蓝染色评价缺损部位产生软骨糖胺聚糖的情况。结果:与B组比较,C组和D组的新生软骨覆盖度、新骨髓的颜色、缺损边缘和表面粗糙度均显著提高(P<0.05);再生软骨的组织学评分软骨表面评分显著改善(P<0.05)。C组与D组具有比其他组更好的基质、细胞分布和表面指数。并且有较厚的透明样软骨组织,具有正常的糖胺聚糖产生。表明该治疗方法可以通过再生透明样软骨且没有不良事件来减少软骨缺陷。结论:工程软骨结合骨碎补治疗可显著改善兔膝关节软骨缺损修复的质量,为临床治疗软骨病变提供重要理论依据。

Objective： To evaluate the effect of rhizome drynaria combined with tissue engineering cartilage on cartilage regenera- tion in experimental rabbits with cartilage defects. Methods： The hlGF-1 gene was transfected into MSCs by using the method of isolation, purification and recombination of transgenic stem cells. The MSCs were transplanted into rabbit bone marrow mesenchymal stem cells （MSCs） in vitro. The cells were further amplified and mixed with acellular dermal matrix （ADM） to construct tissue engineered car- tilage. Twenty-four New Zealand white rabbits, aged 6 months, were randomly divided into 4 groups （A, B, C and D）. six rabbits in each group. Group A and C were transplanted with autologous cartilage. Group B and D were transplanted with modified cells. Group C and D group were fed with 40% Drynaria Decoction, 150ml/d for 4 weeks. Animals were sacrificed at 12 weeks postoperatively, and articular cartilage defects were isolated. Cartilage defect samples were embedded in paraffin blocks and stained with hematoxylin and eosin （H＆E）. Cartilage regeneration was evaluated by gross morphology, including sclerotic shape, color, contour and homogeneity. The quality of regenerated cartilage was assessed by histological scoring. Toluidine blue staining was used to evaluate the occurrence of chondro- genic glycosaminoglycans （GAG）. Results： Compared with group B, the cartilage coverage, the color of new bone marrow, the edge of defect and the surface roughness of group C and D were significantly improved （P〈0.05）; the cartilage surface score of regenerated cartilage was significantly improved P〈0.05）. Groups C and D had better matrix, cell distribution and surface index than the other groups. And had a thick like hyaline cartilage tissue, with the normal glycosaminoglycan production. It is indicated that drynaria combined with tissue engineering cartilage can reduce cartilage defects by regenerating hyaline cartilage. Conclusion： Cartilage combined with drynariae can significantly improve the quality of cartilage defect repair in rabbit knee joint, and provide an important theoretical basis for clinical treatment of cartilage lesions.